| abstract |
This invention relates to the development of recombinant eukaryotic cloning and expression vectors based on unique regulatory elements isolated from autonomously replicating, stable episomal units from human tumor cell lines. More specifically, the unique regulatory elements relate to origins of replication, as well as conferring extrachromosomal stability and maintenance. This cloning and expression vector will accommodate genes that exceed the cosmid limit (greater than 50 kb) and permit their maintenance as autonomously replicating extrachromosomal elements in mammalian cells. Inclusion of telomeres and centromeres would control the replication and segregation and therefore serve as an eventual vehicle for gene replacement therapy. This invention is therefore unique in providing for the expression and autonomous replication of large genes, maintained extrachromosomally, in a vector containing episomal regulatory elements. |